title: The Fc region of an antibody impacts the neutralization of West Nile viruses in different maturation states.
abstract: Flavivirus-infected cells secrete a structurally heterogeneous population of viruses because of an inefficient virion maturation process. Flaviviruses assemble as noninfectious, immature virions composed of trimers of envelope (E) and precursor membrane (prM) protein heterodimers. Cleavage of prM is a required process during virion maturation, although this often remains incomplete for infectious virus particles. Previous work demonstrated that the efficiency of virion maturation could impact antibody neutralization through changes in the accessibility of otherwise cryptic epitopes on the virion. In this study, we show that the neutralization potency of monoclonal antibody (MAb) E33 is sensitive to the maturation state of West Nile virus (WNV), despite its recognition of an accessible epitope, the domain III lateral ridge (DIII-LR). Comprehensive epitope mapping studies with 166 E protein DIII-LR variants revealed that the functional footprint of MAb E33 on the E protein differs subtly from that of the well-characterized DIII-LR MAb E16. Remarkably, aromatic substitutions at E protein residue 306 ablated the maturation state sensitivity of E33 IgG, and the neutralization efficacy of E33 Fab fragments was not affected by changes in the virion maturation state. We propose that E33 IgG binding on mature virions orients the Fc region in a manner that impacts subsequent antibody binding to nearby sites. This Fc-mediated steric constraint is a novel mechanism by which the maturation state of a virion modulates the efficacy of the humoral immune response to flavivirus infection.
authors.affiliation: Viral Pathogenesis Section, Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, NIH, Bethesda, Maryland, USA.
authors.forename: Phong D
authors.initials: PD
authors.lastname: Lee
authors.forename: Swati
authors.initials: S
authors.lastname: Mukherjee
authors.forename: Melissa A
authors.initials: MA
authors.lastname: Edeling
authors.forename: Kimberly A
authors.initials: KA
authors.lastname: Dowd
authors.forename: S Kyle
authors.initials: SK
authors.lastname: Austin
authors.forename: Carolyn J
authors.initials: CJ
authors.lastname: Manhart
authors.forename: Michael S
authors.initials: MS
authors.lastname: Diamond
authors.forename: Daved H
authors.initials: DH
authors.lastname: Fremont
authors.forename: Theodore C
authors.initials: TC
authors.lastname: Pierson
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology; D057134:Antibodies, Neutralizing / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology*; D018604:Epitope Mapping; D057809:HEK293 Cells; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000276:immunology*; D009500:Neutralization Tests; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D014901:West Nile Fever / Q000276:immunology / Q000821:virology*; D014902:West Nile virus / Q000737:chemistry / Q000235:genetics / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D007141:Immunoglobulin Fc Fragments; D014759:Viral Envelope Proteins